叶酸途径调控对谷氨酸棒杆菌syps062积累l丝氨酸的影响-effects of folic acid pathway regulation on l - serine accumulation in corynebacterium glutamicum syps 062.docxVIP

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叶酸途径调控对谷氨酸棒杆菌syps062积累l丝氨酸的影响-effects of folic acid pathway regulation on l - serine accumulation in corynebacterium glutamicum syps 062.docx

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叶酸途径调控对谷氨酸棒杆菌syps062积累l丝氨酸的影响-effects of folic acid pathway regulation on l - serine accumulation in corynebacterium glutamicum syps 062

II Ea--Abstract Abstract Ea - - ηle tetrahydrofolate (THF) was generated in folate pathway in C. glutamicum ，and engaged in regulating SHMT enzyme. SHMT enz归ne catalysed the conversion of L-serine into glycine. The study focused on a L-serine producing strain 巳 glutamicum SYPS-062， which can accumulate L-serine directly from sugar substances. The gene pabAB overexpression strain C.glutamicum SYPS062(ppabAB) and the gene pabAB deletion strain lutamicum SYPS-0626pabAB were constructed ，the effect of pabAB ωthe L-serine accumulation in C. glutlαmicum SYPS062，and the effect of other substances in the folate pathway to the L serine acumulation were also studied. The results wereωfollowing: 刊e homologous 仕agment 6pabAB w部 amplified through crossover PCR， and was ligased to pK 18mobsacB ，then the 巳 glu阳micum SYPS-0626pabAB wωconstructed through homologous recombination. 。T)he results showed th副 the ADC synthase raised 33% with the gene pabAB overexpression compared with the original strain，while it decreased 61% after the gene pabAB was deleted. Meanwhile the SHMT activity of C. glutamicum SYPS-062 (ppabAB) increased 30%，the μm raised 48%，and the Yp/x decreased 36.2% compared with that of the original srtain C. glutamicum SYPS-062; The SHMT activity of C. glutamicum SYPS-0626pαbAB decresaed 20% when compared to that of the original strain，the Pm decreased 32%，while the Yp/x increased 12% compared with the original strain. (3) The accumulation of L-serine by 巳 gluωmicum SYPS-0626pαbAB could reach up ω7.8 gIL in the optimized medium，which increased 14% compared to that of the original strain. Key words: folate pathway，pabAB ，overexpression ，homologuous recombinatio n， gene deletion，fermentation optimization 2.1.1 材料……………………………………… 2.1.1 材料……………………………………… 9 2.1.2 实验方法……·…………………………… 11 2.2 结果与讨论………………………… 17 2.2.1 向源片段f.:::.pαbAB 的构建111...………………………………..……………………17 2.2.2 重组质粒 pK18mobsacB-6pabAB 的构建及验